rat ly6c antibody  (Bio-Rad)

Journal: JCI Insight

Article Title: The CHI3L1-neutrophil axis drives immune suppression and breast cancer metastatic dissemination

doi: 10.1172/jci.insight.199307

Figure Lengend Snippet: ( A ) Staining of WT, Chi3l1 OE, Stat3 –/– , and Stat3 –/– Chi3l1 OE MIC mammary glands at 2 weeks after induction for CD8, granzyme B (GZMB), CD3, Pan-CK, and DAPI. ( B ) RNA FISH against IFN-γ with staining for CD3, Pan-CK, and DAPI on mammary tissue from WT, Chi3l1 OE, Stat3 –/– , and Stat3 –/– Chi3l1 OE MIC mammary glands at 2 weeks after induction. ( C – G ) Quantification of total CD3 + CD8 + , CD3 + CD8 + GZMB + , and tumor-infiltrating CD3 + CD8 + T cells, CD3 + IFN-γ + cells, and Ly6G + cells in WT ( n = 5), Chi3l1 OE ( n = 8), Stat3 –/– ( n = 6), and Stat3 –/– Chi3l1 OE ( n = 9) MIC mammary glands at 2 weeks after induction. ( H ) Staining of WT, Chi3l1 OE, Stat3 –/– , and Stat3 –/– Chi3l1 OE MIC mammary glands at 2 weeks after induction using antibodies against Ly6G, MPO, NE, pan-CK, and DAPI. ( I and J ) Quantification of total Ly6G + MPO + cells and Ly6G + MPO + NE + cells in WT ( n = 5), Chi3l1 OE ( n = 8), Stat3 –/– ( n = 6), and Stat3 –/– Chi3l1 OE ( n = 9) MIC mammary glands at 2 weeks after induction. ( K ) Quantification of CitH3 immunoblots normalized to α-tubulin. ( L ) Immunoblots for CitH3 and α-tubulin on WT ( n = 3), Chi3l1 OE ( n = 3), Stat3 –/– ( n = 3), and Stat3 –/– Chi3l1 OE ( n = 3) MIC mammary glands at 2 weeks after induction. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by 1-way ANOVA with Tukey’s post hoc test. Scale bars: 100 μm.

Article Snippet: To deplete neutrophils, cohorts of MTB MIC CHI3L1 OE mice were randomized to receive 400 μg of purified anti-LY6G rat antibody (1A8, Bio X Cell BE0075-1) or 400 μg of IgG2a rat isotype control (2A3, Bio X Cell BE0089) via intraperitoneal (i.p.) injections twice weekly.

Techniques: Staining, Western Blot

Journal: JCI Insight

Article Title: The CHI3L1-neutrophil axis drives immune suppression and breast cancer metastatic dissemination

doi: 10.1172/jci.insight.199307

Figure Lengend Snippet: ( A ) H&E staining of mammary glands from IgG2a- and anti-Ly6G–treated MIC Stat3 –/– Chi3l1 OE mice at 2 weeks after induction. ( B ) Quantification of hyperplastic area in IgG2a-treated ( n = 8) and anti-Ly6G–treated ( n = 8) MIC Stat3 –/– Chi3l1 OE mice. Represented as percentage of mammary gland area. ( C ) Staining of mammary tissue from MIC Stat3 –/– Chi3l1 OE mice treated with anti-Ly6G or IgG2a for 2 weeks, for GZMB, CD8, CD3, pan-CK, and DAPI. ( D – F ) Quantification of CD3 + CD8 + , CD3 + CD8 + GZMB + , and tumor-infiltrating CD3 + CD8 + T cells in IgG2a-treated ( n = 8) and anti-Ly6G–treated ( n = 8) MIC Stat3 –/– Chi3l1 OE mammary glands. ( G ) Staining of mammary tissue from MIC Stat3 –/– Chi3l1 OE mice treated with anti-Ly6G or IgG2a, for PD-1, CD4, CD3, pan-CK, and DAPI. ( H – J ) Quantification of CD3 + CD4 + , CD3 + CD4 + PD-1 + , and tumor-infiltrating CD3 + CD4 + T cells in IgG2a-treated ( n = 8) and anti-Ly6G–treated ( n = 8) MIC Stat3 –/– Chi3l1 OE mammary glands. ( K ) H&E staining of mammary glands from IgG2a- and anti-Ly6G–treated MIC Chi3l1 OE mice at 2 weeks after induction. ( L ) Quantification of hyperplastic area in IgG2a-treated ( n = 6) and anti-Ly6G–treated ( n = 8) MIC Chi3l1 OE mice. Represented as percentage of mammary gland area. ( M ) Staining of mammary tissue from MIC Chi3l1 OE mice treated with anti-Ly6G or IgG2a for 2 weeks, for GZMB, CD8, CD3, pan-CK, and DAPI. ( N – P ) Quantification of CD3 + CD8 + , CD3 + CD8 + GZMB + , and tumor-infiltrating CD3 + CD8 + T cells in IgG2a-treated ( n = 6) and anti-Ly6G–treated ( n = 8) MIC Chi3l1 OE mammary glands. ( Q ) Staining of mammary tissue from MIC Chi3l1 OE mice treated with anti-Ly6G or IgG2a for 2 weeks, for PD-1, CD4, CD3, pan-CK, and DAPI. ( R – T ) Quantification of CD3 + CD4 + , CD3 + CD4 + PD1 + , and tumor-infiltrating CD3 + CD4 + T cells in IgG2a-treated ( n = 6) and anti-Ly6G–treated ( n = 8) MIC Chi3l1 OE mammary glands. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by unpaired Student’s t test. Scale bars: 100 μm in A , C , G , M , and Q ; 500 μm in K .

Article Snippet: To deplete neutrophils, cohorts of MTB MIC CHI3L1 OE mice were randomized to receive 400 μg of purified anti-LY6G rat antibody (1A8, Bio X Cell BE0075-1) or 400 μg of IgG2a rat isotype control (2A3, Bio X Cell BE0089) via intraperitoneal (i.p.) injections twice weekly.

Techniques: Staining

Journal: JCI Insight

Article Title: The CHI3L1-neutrophil axis drives immune suppression and breast cancer metastatic dissemination

doi: 10.1172/jci.insight.199307

Figure Lengend Snippet: ( A ) H&E staining of WT, Chi3l1 OE, Stat3 –/– , and Stat3 –/– Chi3l1 OE MIC lungs at mammary tumor endpoint. ( B ) Percentage of WT ( n = 12), Chi3l1 OE ( n = 8), Stat3 –/– ( n = 13), and Stat3 –/– Chi3l1 OE ( n = 11) MIC mice with pulmonary metastases. ( C ) Quantification of metastatic area (percent total lung area) in WT ( n = 12), Chi3l1 OE ( n = 8), Stat3 –/– ( n = 13), and Stat3 –/– Chi3l1 OE ( n = 11) MIC lungs. ( D ) Quantification of PyMT + cells (percent live cells in blood) of WT ( n = 4) and Chi3l1 OE ( n = 3) MIC mice. ( E ) FACS for circulating PyMT + cells in WT and Chi3l1 OE MIC blood at 2 weeks after induction. ( F ) Staining of WT and Chi3l1 OE MIC mammary glands at 2 weeks after induction for laminin, collagen IV, pan-CK, and DAPI. ( G and H ) Quantification of area occupied by laminin or collagen IV (percent total mammary gland area) in WT ( n = 7) and Chi3l1 OE ( n = 8) MIC mammary glands. ( I ) FACS for circulating PyMT + cells in blood of Chi3l1 OE MIC mice after anti-Ly6G or IgG2a treatment for 2 weeks. ( J ) Quantification of PyMT + cells (percent live cells in the blood) of Chi3l1 OE MIC mice treated with IgG2a ( n = 6) or anti-Ly6G ( n = 5). ( K ) Staining of mammary tissue from MIC Chi3l1 OE mice treated with anti-Ly6G or IgG2a for laminin, collagen IV, pan-CK, and DAPI. ( L and M ) Quantification of area occupied by laminin and collagen IV (percent mammary gland area) in IgG2a-treated ( n = 6) and anti-Ly6G–treated ( n = 8) MIC Chi3l1 OE mammary glands. ( N ) Schematic of Transwell invasion assay through synthetic ECM. Created in BioRender (Muller W, 2026, https://BioRender.com/4lhz2a3 ). ( O ) ECM-invading PyMT + cells with PBS, rmChi3l1, or murine neutrophils, stained with crystal violet. Neutrophils + PBS (no PyMT cells) is shown as negative control. ( P ) Quantification of invading PyMT + cells. n = 3 technical replicates for each condition. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by 1-way ANOVA with Tukey’s post hoc test ( C and P ) or by unpaired Student’s t test (for D , G , H , J , L , and M ). Scale bars: 1 cm in A , left panels; 1 mm in A , right panels; 100 μm in F , K , and O .

Article Snippet: To deplete neutrophils, cohorts of MTB MIC CHI3L1 OE mice were randomized to receive 400 μg of purified anti-LY6G rat antibody (1A8, Bio X Cell BE0075-1) or 400 μg of IgG2a rat isotype control (2A3, Bio X Cell BE0089) via intraperitoneal (i.p.) injections twice weekly.

Techniques: Staining, Transwell Invasion Assay, Negative Control

Journal: JCI Insight

Article Title: The CHI3L1-neutrophil axis drives immune suppression and breast cancer metastatic dissemination

doi: 10.1172/jci.insight.199307

Figure Lengend Snippet: ( A ) Immunoblots for Chi3l1, Stat3, and α-tubulin on WT ( n = 7) and Chi3l1 –/– ( n = 6) MIC mammary endpoint tumors. ( B ) Representative H&E staining of WT and Chi3l1 –/– MIC lungs at mammary tumor endpoint. ( C ) Percentage of WT ( n = 11) and Chi3l1 –/– ( n = 11) MIC mice with pulmonary metastases. ( D ) Quantification of total pulmonary metastatic area in WT ( n = 11) and Chi3l1 –/– ( n = 11) MIC lungs. Represented as percentage of total lung area. ( E ) Number of lung metastatic lesions in WT ( n = 11) and Chi3l1 –/– ( n = 11) MIC lungs. ( F ) Staining of WT and Chi3l1 –/– MIC mammary glands at 2 weeks after induction, for Ly6G, MPO, NE, pan-CK, and DAPI. ( G and H ) Quantification of Ly6G + cells and Ly6G + MPO + NE + cells in WT ( n = 7) and Chi3l1 –/– ( n = 10) MIC mammary glands at 2 weeks after induction. ( I ) Staining of WT and Chi3l1 –/– MIC mammary glands at 2 weeks after induction, for laminin, collagen IV, pan-CK, and DAPI. ( J and K ) Quantification of area occupied by laminin or collagen IV in WT ( n = 7) and Chi3l1 –/– ( n = 10) MIC mammary glands at 2 weeks after induction. Represented as percentage of total mammary gland area. ( L ) FACS sorting for PyMT + tumor cells in the blood of WT MIC mice treated with anti-Chi3l1 neutralizing antibody or IgG2b isotype control at 2 weeks after induction. ( M ) Quantification of PyMT + cells as percentage of total live cells in the blood of WT MIC mice treated with IgG2b ( n = 5) or anti-Chi3l1 ( n = 5) for 2 weeks. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by unpaired Student’s t test. Scale bars: 1 mm in B , left panels; 100 μm in B , right panels; 10 μm in F and I .

Article Snippet: To deplete neutrophils, cohorts of MTB MIC CHI3L1 OE mice were randomized to receive 400 μg of purified anti-LY6G rat antibody (1A8, Bio X Cell BE0075-1) or 400 μg of IgG2a rat isotype control (2A3, Bio X Cell BE0089) via intraperitoneal (i.p.) injections twice weekly.

Techniques: Western Blot, Staining, Control

Journal: Scientific Reports

Article Title: Constitutive interaction between neutrophils and von Willebrand factor in peripheral blood

doi: 10.1038/s41598-025-10321-6

Figure Lengend Snippet: Interaction between mouse peripheral neutrophils and VWF. (A) Representative flow cytometry histogram illustrating VWF presence on the surface of neutrophils from wildtype mice. (B) Statistical analysis of mean fluorescence intensity (MFI) for VWF on neutrophil surfaces relative to control ( n = 11–12). (C) SDS-agarose gel electrophoresis comparing VWF multimer in neutrophils (Ly6G + ) versus other leukocytes isolated from wildtype mouse peripheral blood. Full-length gel is included in supplementary figure S5A. (D) Flow cytometry scatter plot displaying leukocyte populations from GFP-VWF transgenic mice stained with anti-Ly6G antibody. (E) Overlay of flow cytometry histograms comparing VWF signal intensity between platelets and neutrophils in GFP-VWF transgenic mice. (F) Statistical summary ( n = 4) corresponding to data in preceding panels. Statistical significance was assessed using t-tests for pairwise comparisons and one-way ANOVA for multiple groups, with significance denoted as **** p < 0.0001, * p < 0.05.

Article Snippet: Rat anti-mouse Ly6G antibody , Bio X cell , Cat #BE0075-1.

Techniques: Flow Cytometry, Fluorescence, Control, Agarose Gel Electrophoresis, Isolation, Transgenic Assay, Staining

Journal: Scientific Reports

Article Title: Constitutive interaction between neutrophils and von Willebrand factor in peripheral blood

doi: 10.1038/s41598-025-10321-6

Figure Lengend Snippet: The neutrophil-VWF association is absent in bone marrow. Representative confocal fluorescence microscopy images of bone marrow from GFP-VWF transgenic mice, showing intrinsic GFP-VWF fluorescence in green, anti-mouse Ly6G staining labeling neutrophils in red, and nuclei stained with DAPI in blue. Scale bars represent magnifications of 100 μm and 10 μm, respectively.

Article Snippet: Rat anti-mouse Ly6G antibody , Bio X cell , Cat #BE0075-1.

Techniques: Fluorescence, Microscopy, Transgenic Assay, Staining, Labeling

Journal: Scientific Reports

Article Title: Constitutive interaction between neutrophils and von Willebrand factor in peripheral blood

doi: 10.1038/s41598-025-10321-6

Figure Lengend Snippet: Depletion of neutrophils increases the VWF level in plasma. (A) Representative flow cytometry profiles showing the effect of neutrophil depletion via anti-Ly6G antibody. The IgG isotype serves as control. (B) SDS-agarose gel electrophoresis image showing VWF multimers before and after neutrophil depletion. Full-length gel is included in supplementary figure S5B. The averaged fold change in plasma VWF levels in mice post-neutrophil depletion compared to control is shown on the right ( n = 6). (C) Quantitative assessment of plasma VWF propeptide (VWFpp) levels in mice following anti-Ly6G antibody injection at 0 and 12 h, with comparative analysis of VWFpp changes between time points and control groups, as measured by immunoblotting. (D) Comparison of VWF mRNA levels in lung and aorta tissues of anti-Ly6G antibody-treated versus control IgG-treated mice ( n = 6, Left) and statistical analysis of mRNA levels of liver genes ( asgr1 , asgr2 , lrp1 , sra1 , stab2 , mgl1 , mgl2 ) in the same groups ( n = 6, Right). Statistical analyses were performed using t-tests for pairwise comparisons, with significance denoted as* p < 0.05, and “n.s.” for not significant.

Article Snippet: Rat anti-mouse Ly6G antibody , Bio X cell , Cat #BE0075-1.

Techniques: Clinical Proteomics, Flow Cytometry, Control, Agarose Gel Electrophoresis, Injection, Western Blot, Comparison